Documenti di Didattica
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74, 1968] 55
By A. D. Portno
{Brewing Industry Research Foundation, Nutficld, Surrey)
Results from a wide range of systems for the continuous fermentation of brewer's
wort demonstrate that two criteria determine the operational aspects of such
systems. These are:
1I) The degree of mixing, and
(2) The degree of restraint over the escape of yeast.
A reduction in the ability of yeast to ferment occurs when high concentrations of
yeast are maintained and is evident to a greater extent in homogeneous systems than
in those which maintain a fermentation gradient.
Results suggest that satisfactory systems must allow partial escape of yeast and
that a minimum degree of heterogeneity should be maintained. These requirements
necessitate a system which either maintains a concentration gradient in an elongated
vessel or comprises a series of homogeneous vessels with a feed-back of cells so that,
in each instance, yeast in the system is regularly exposed to a high concentration of
nutrients.
under gravity in order to build up high it has an internal diameter of f in. and is 35 in.
concentrations.6'10-11 •28 Under such condi long. The vessel broadens at its upper end
tions the fermentative capacity of the yeast to form a glass bulb which serves as a cham
decreases and this is reflected by an increase ber within which foam is confined. Fer
in the specific gravity of the liquid within menting liquid which contains yeast is
the system; this may induce the yeast to withdrawn from the lower end of the tube
deflocculate and therefore lead to it being and recirculated to the foaming chamber
washed out of the apparatus. Such an thus ensuring complete mixing throughout
effect has been observed both in these the system. The tubular portion of the
laboratories and elsewhere.10'11 fermenting vessel is enclosed within a glass
In order to allow such a sequence of events tube into which wort is introduced.
to proceed beyond this point and thus to Equilibration occurs across the filter-tube
facilitate examination of the process in as the liquid is induced to flow alongside it,
greater detail, a new system of continuous causing wort to enter the fermentation
fermentation was constructed and is illus circuit and the products of fermentation to
trated in Fig. 1. The apparatus employs a leave: the nitration properties of the tube
are such as to prevent completely the escape
Arphon of j'east cells. Trials showed that free
Syphoning Foaming
break hamber transfer of liquid across the filter-tube was
.eve. ,—-&fi«L insufficient to prevent the deposition of a
1
Level
layer of yeast on the internal surface and
therefore, in order to prevent this, an
automatic back-flushing system was attached
fluctuation to the liquid outlet. This, by an intermittent
syphoning action, induced a reciprocating
transfer of liquid across the filter-tube and
thus ensured freedom from obstruction.
Fig. 2 shows the changes in specific gravity
and yeast viability, as estimated from
methylene-blue staining, which occurred
during a fermentation when a powdery yeast
(N.C.Y.C. No. 240) was used. During this
trial no yeast was allowed to escape and the
system was thus operating under conditions
when the closure index was 100%; it can be
seen that under these conditions the specific
gravity of the liquid in the system fluctuated
Wort
considerably during a distinct overall in
crease. These altered levels of specific
Yeast
circulation
gravity were reflected by changes in the systems are employed, steady-state condi
ability of the yeast to reduce methylene tions become increasingly difficult to achieve
blue. as the attenuation limit of the wort is
Examination of the sugars present in the approached.
medium during the course of the trial Changes which occurred in the specific
showed that the fluctuations in specific gravity of the contents of an open-homo
gravity were largely caused by altered levels geneous continuous fermentation system
of maltose in the fermenting medium; this is which was operated for four weeks at an
plainly a reflection of changes in the ability efficiency of 0-8 and under constant condi
of the yeast to remove it. Table I shows tions are shown in Fig. 3. It may be seen
that a rather regular pattern of changes
TABLE I occurred similar to those which were observed
Changes in Specific Gravity and Concentra in the closed fermentor which was described
tions of Maltose which Occurred in a Closed
earlier, although in the present example the
System of Continuous Fermentation
magnitude of the fluctuations was reduced
and no overall decrease in the fermentative
Concentration
capacity of the yeast was observed.
Time Specific of maltose
Sample* (days) 1 gravity (g. per litre)
A 4 10063 10
B 0 10122 120
C 12 10070 20
D 18 10135 17-0
E 30 1-0176 21-0
Wort — 10400 430 16
Time (days)
* Letters A-E refer to points labelled in Fig. 2. Fig. 3.—Measurements of specific gravity during the
operation of an open-homogeneous continuous
alterations in concentrations of maltose tormentor.
producing partially attenuated liquid of minimize the effect and therefore to prevent
specific gravity 1-035 from brewer's wort of wash-out, but is insufficient to suppress all
specific gravity 1-040. instability.
In the second investigation, the same In the third trial the two fermentation
simple continuous fermentors were employed systems were again operated side by side;
and were operated under identical conditions, on this occasion the pre-treatment vessel
using the same wort and yeast as in the was absent and the systems thus were
former trial. The two fermentation systems identical in all respects. The specific gravity
differed only in that an additional stirred of the contents of both vessels rose to
vessel was installed in series with one of between 1-014 and 1-017 and at this point
them, yeast from the main fermenting vessel the inflow of wort was stopped. Into one
being recirculated to meet the wort which fermentor 100 ml. of a solution of maltose
entered through the small pre-treatment was added; this contained 51-6 g. of maltose
vessel. This arrangement was intended to and resulted in the total concentration of this
ensure that yeast which was recirculated was sugar in the vessel being elevated to that of
exposed to a high concentration of nutrients unfermented wort of specific gravity 1*040
in order to stimulate induction of the neces (43 g. per litre). 100 ml. of sterile water was
sary enzyme systems. The specific gravity in introduced into the other fermentor as a
the main fermenting vessels was 1-011 and control.
that in the pre-treatment vessel 1-026. The Fig. 6 shows the results obtained when,
volumes of liquid in the vessels was 1200 ml. after allowing the fermentors to stand for
in the main fermenting vessel and 600 ml. in 24 hr., the introduction of wort was recom
the pre-treatment vessel. Both fermentation menced at the former rate. In the control,
systems were operated at an efficiency of 3 to which water had been added, the specific
which corresponds to a dilution rate of gravity continued to rise at an increasing
0-125 h.-i rate and ultimately induced deflocculation
Fig. 5 shows the changes in specific gravity of the yeast and wash-out. On the other
which were observed when the systems were hand, exposure of the yeast to the high
operated for a period of 20 days. In the concentration of maltose delayed the sub
case of the single-vessel fermentation system, sequent rise in specific gravity. However,
the specific gravity rose after 10 days by reference to the first trial and to Fig. 5 it
resulting in wash-out and the subsequent is clear that maltose alone was less effective
functioning of the system as an open con than wort in so doing and therefore, although
tinuous fermentor attenuating through 4 concentrations of maltose which influence the
degrees only. A parallel rise in specific ability of yeast to remove and ferment it are
gravity was observed in the two-vessel of obvious importance in this respect, other
system although its magnitude was con
siderably reduced and the fermentative
capacity of the yeast recovered before the
point was reached where deflocculation
i-ok
would have occurred; this allowed recovery
and a consequent reduction in the specific
gravity to the original level. Plainly this
degree of heterogeneity is sufficient to
Time (days)
20
Fig. 6.—The effect on specific gravity of the addition
Time (days)
of maltose to a closed-homogeneous system as
Fig. 6.—Changes in specific gravity in two systems of the fermentative ability of the yeast com
continuous fermentation:— menced to decline (O—O—Q) and results
(a) A single-vessel system (0—0—0) and obtained from a control to which water was
(6) A two-vessel system (■—■—■). added (©—#—#).
60 PORTNO: CONTINUOUS FERMENTATION [J. Inst. Brew.
sugars, and possibly additional materials of The growth of yeast in continuous fer
the type which are required for the synthesis mentation systems is frequently considered
of enzymes, are probably involved. as specific growth rate (p). This expression
It is possible that, as a yeast cell ages, its is a measure of the rate at which new cells
ability to bring about fennentation declines are produced and is related to the generation
and that, at a point dependent upon fermen or doubling time.
tation conditions and the individual strain However, the specific growth rate is
of yeast, its fermentative capacity becomes essentially a measure of the rate at which
seriously impaired. Clearly, therefore, it new cells are produced and, as such, was
seems desirable to adjust the conditions devised in connection with studies on the
under which a continuous fermentor operates continuous culture of micro-organisms when
so that the time during which yeast cells the main objective was to produce such
remain in the system is regulated. This organisms at or near maximum rates.
would enable the turnover to be maintained In the continuous fermentation of wort the
at such a level that a large proportion of the end product is the fermented liquor and
ageing cells would leave the system before growth of yeast is clearly a secondary con
the time at which their fermentative sideration. In these circumstances such
capacities commenced to decline. growth is required for two purposes:—
Fig. 7 shows the rate of reduction in the (a) to produce new cells at a rate at
ability of cells to reduce methylene blue with least equal to the rate at which
time in the case of two closed systems of
(i) cells escape from the fennentation
continuous fermentation, one of which was
system,
homogeneous whilst the other was hetero
(ii) old cells lose the ability to
geneous. It can be seen by reference to the
ferment; and
Figure how the fluctuations which occur in a
homogeneous system confuse the secondary (b) to regulate the flavour of the finished
effect which is predominantly due to an beer which, in several respects, has
excessive closure-index. Such fluctuations been shown to be influenced by the
do not occur in heterogeneous continuous amount of yeast growth which
fermentors and although the mean slope of occurs.18'18
the curve which depicts the reduction in In order that a continuous fermentation
viability is the same in both types of system system should function under steady con
this becomes confused in the case of homo ditions, it is plainly essential to maintain a
geneous fermentors by the fluctuations constant concentration of active yeast
which occur around the mean. In this within it and the population level which is
instance a mean reduction of 40% in the necessary will depend on both the efficiency
ability of yeast cells to reduce methylene at which the system is required to operate
blue occurred over a period of 40 days and and the fermentative ability of the par
at closure indices of 94% and 100%. ticular yeast involved.
Fig. 8 shows the concentrations of yeast
required to ferment wort from a specific
gravity of 1-040 to 1-010 over a range of
efficiencies from 1 to 13 and is based on the
results reported by several authors8'8'9'8'10-13
all of whom claimed that the continuous
fennentation systems from which the results
were obtained operated under steady condi
tions. Clearly, if the fermentative capacity
of the yeast is maintained at a constant level
20 30 40 SO these proportions should stand in linear
Time (days) relationship. However, in the presence of
Fig. 7.—Alterations in the ability of cells to reduce very high concentrations of yeast, the volume
methyleno blue which occurred in occupied by the yeast cells in the fermentation
(a) a closed-homogeneous continuous fer
vessels becomes a significant proportion of
mentor (O—D—D)
and (6) a closed-heterogeneous continuous fer the total capacity and extrapolation of the
mentor (O—O—O). graph is thus only possible if compensation
Vol. 74, 1968] PORTNO: CONTINUOUS FERMENTATION 61